Rhodopseudomonas palustris collagen-like recombinant protein purification using an aqueous two-phase system
The potential use of recombinant collagen-like protein (recCLP) extracted from bacteria as disease-free collagen has been studied over the past decade. However, the complexity of the downstream processing generates high demand for an efficient and low-cost purification method. Aqueous two-phase syst...
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International Islamic University Malaysia-IIUM
2023
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| Online Access: | http://umpir.ump.edu.my/id/eprint/40637/ http://umpir.ump.edu.my/id/eprint/40637/1/Rhodopseudomonas%20palustris%20collagen-like%20recombinant.pdf |
| _version_ | 1848826104857493504 |
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| author | Nursyahidatul Azwa, Awang Azura, Amid Zatul Iffah, Mohd Arshad |
| author_facet | Nursyahidatul Azwa, Awang Azura, Amid Zatul Iffah, Mohd Arshad |
| author_sort | Nursyahidatul Azwa, Awang |
| building | UMP Institutional Repository |
| collection | Online Access |
| description | The potential use of recombinant collagen-like protein (recCLP) extracted from bacteria as disease-free collagen has been studied over the past decade. However, the complexity of the downstream processing generates high demand for an efficient and low-cost purification method. Aqueous two-phase system (ATPS) was adopted as a new approach to the recovery of biomolecules due to its simple, benign, and straightforward process. This study aimed to purify recombinant collagen-like protein from Rhodopseudomonas palustris using ATPS formed by a polymer/salt system. Recombinant collagen-like protein from R. palustris was partitioned in ATPS composed of polyethylene glycol (PEG) and potassium phosphate and several factors that influence the protein partitioning such as volume ratio, system pH, the concentration of polymer and salt were studied. Then, optimization of the selected ATPS conditions (PEG and salt concentration) were performed using response surface methodology (RSM). Results showed that the optimum conditions were found in ATPS with 24.80% (w/w) PEG 2000 and 29.23% (w/w) potassium phosphate with recCLP concentration of 3.23 ± 0.12 mg/mL with purification factor 7.48 ± 0.3. In comparison with the affinity chromatography method, ATPS was found to be low-cost, and time-saving with a higher protein recovery. Hence, this study demonstrated the potential application of ATPS in the recovery of recombinant CLPs for large-scale downstream processing |
| first_indexed | 2025-11-15T03:39:31Z |
| format | Article |
| id | ump-40637 |
| institution | Universiti Malaysia Pahang |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T03:39:31Z |
| publishDate | 2023 |
| publisher | International Islamic University Malaysia-IIUM |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | ump-406372024-04-30T06:39:11Z http://umpir.ump.edu.my/id/eprint/40637/ Rhodopseudomonas palustris collagen-like recombinant protein purification using an aqueous two-phase system Nursyahidatul Azwa, Awang Azura, Amid Zatul Iffah, Mohd Arshad QD Chemistry T Technology (General) TA Engineering (General). Civil engineering (General) TP Chemical technology The potential use of recombinant collagen-like protein (recCLP) extracted from bacteria as disease-free collagen has been studied over the past decade. However, the complexity of the downstream processing generates high demand for an efficient and low-cost purification method. Aqueous two-phase system (ATPS) was adopted as a new approach to the recovery of biomolecules due to its simple, benign, and straightforward process. This study aimed to purify recombinant collagen-like protein from Rhodopseudomonas palustris using ATPS formed by a polymer/salt system. Recombinant collagen-like protein from R. palustris was partitioned in ATPS composed of polyethylene glycol (PEG) and potassium phosphate and several factors that influence the protein partitioning such as volume ratio, system pH, the concentration of polymer and salt were studied. Then, optimization of the selected ATPS conditions (PEG and salt concentration) were performed using response surface methodology (RSM). Results showed that the optimum conditions were found in ATPS with 24.80% (w/w) PEG 2000 and 29.23% (w/w) potassium phosphate with recCLP concentration of 3.23 ± 0.12 mg/mL with purification factor 7.48 ± 0.3. In comparison with the affinity chromatography method, ATPS was found to be low-cost, and time-saving with a higher protein recovery. Hence, this study demonstrated the potential application of ATPS in the recovery of recombinant CLPs for large-scale downstream processing International Islamic University Malaysia-IIUM 2023 Article PeerReviewed pdf en cc_by_nc_4 http://umpir.ump.edu.my/id/eprint/40637/1/Rhodopseudomonas%20palustris%20collagen-like%20recombinant.pdf Nursyahidatul Azwa, Awang and Azura, Amid and Zatul Iffah, Mohd Arshad (2023) Rhodopseudomonas palustris collagen-like recombinant protein purification using an aqueous two-phase system. IIUM Engineering Journal, 24 (1). pp. 40-56. ISSN 1511-788X. (Published) https://doi.org/10.31436/iiumej.v24i1.2468 https://doi.org/10.31436/iiumej.v24i1.2468 |
| spellingShingle | QD Chemistry T Technology (General) TA Engineering (General). Civil engineering (General) TP Chemical technology Nursyahidatul Azwa, Awang Azura, Amid Zatul Iffah, Mohd Arshad Rhodopseudomonas palustris collagen-like recombinant protein purification using an aqueous two-phase system |
| title | Rhodopseudomonas palustris collagen-like recombinant protein purification using an aqueous two-phase system |
| title_full | Rhodopseudomonas palustris collagen-like recombinant protein purification using an aqueous two-phase system |
| title_fullStr | Rhodopseudomonas palustris collagen-like recombinant protein purification using an aqueous two-phase system |
| title_full_unstemmed | Rhodopseudomonas palustris collagen-like recombinant protein purification using an aqueous two-phase system |
| title_short | Rhodopseudomonas palustris collagen-like recombinant protein purification using an aqueous two-phase system |
| title_sort | rhodopseudomonas palustris collagen-like recombinant protein purification using an aqueous two-phase system |
| topic | QD Chemistry T Technology (General) TA Engineering (General). Civil engineering (General) TP Chemical technology |
| url | http://umpir.ump.edu.my/id/eprint/40637/ http://umpir.ump.edu.my/id/eprint/40637/ http://umpir.ump.edu.my/id/eprint/40637/ http://umpir.ump.edu.my/id/eprint/40637/1/Rhodopseudomonas%20palustris%20collagen-like%20recombinant.pdf |